DE60118714T2 - SOLID PHASE SYNTHESIS AND REAGENTS THEREFOR - Google Patents

Abstract

A reagent composition adapted to be coated onto a support surface in order to provide that surface with a high density of reactive groups. The surface, thus coated, can be used for any suitable purpose, and is particularly well suited for use as a solid phase synthesis support surface. The synthesis support surface, in turn, can be used in repetitive and combinatorial syntheses such as the synthesis of polynucleotides, polypeptides and polysaccharides. The polymeric coating can be used to provide increased effective surface area, particularly in situations in which the effective area of the support surface is itself limited, as on the surface of a silicon wafer. In so doing, the polymeric coating provides an optimal combination of such properties as reactive density and surface area or volume.

Description

TECHNICAL TERRITORY

In In one aspect, the present invention relates to methods, Reagents and carrier surfaces for use in solid phase synthesis (e.g., repetitive or combinatorial). In another aspect, the invention relates to substituted ones Polyacrylamidreagentien. In yet another appearance concerns The Invention Reagents for Use in the Modification of Carrier surfaces and in particular the use of photochemical means to such Reagents to such surfaces to add.

BACKGROUND THE INVENTION

Solid phase synthesis has been since the seminal work of R.B. Merrifield in the year Enormously developed in 1963. Typically, the reactions used the same as an ordinary one Synthesis in which one of the reactants is anchored to a solid support becomes. A solid-phase synthesis, for example, for the synthesis used by polynucleotides, polysaccharides and polypeptides be, as well for other applications in repetitive syntheses and combinatorial Chemistry.

Of the The main advantage of the solid phase process is that the support (including all attached to this Reagents) insoluble remains and therefore easily separated from all other reagents becomes. Excess reagents, other reaction products and by-products become fast and efficient when removing the solvents away. Purification of the solid phase species is fast and complete, as well the entire process can be automated.

In In recent years, the principles of solid phase synthesis are on a new methodology has been applied, called "combinatorial Chemistry "known is. Scientists use combinatorial chemistry to large populations of molecules, or collections that are efficiently screened en masse can. By making larger, more diversified Connection collections can Companies increase the likelihood that they will make new connections from considerable find therapeutic and commercial value. The area presents an approach of chemistry and biology, through fundamental advances in miniaturization, roboter technology and receptor development possible is.

As for the conventional Drug development, combinatorial chemistry is based on methodologies organic synthesis. The difference is the scope - instead the synthesis of a single compound uses the combinatorial Chemistry an automation and miniaturization to large collections of synthesizing compounds. However, there are large collections no active compounds independent manufacture, must Scientists also have the active components within this find enormous populations. Therefore, combinatorial organic Synthesis not random, but systematically and repetitively, using sets of chemical "education blocks" to produce various sets of to form molecular units.

It There are at least three usual ones approaches for combinatorial organic synthesis. During a arranged, spatially addressable Synthesis become educational blocks systematically implemented in individual reaction wells or positions, to separate "discrete Molecules. "Active Connections are identified by their location on the grid. This process is in great scale (as in the Parke-Davis Pharmaceutical "DIVERSOMER" method) and as well in miniature (as in the Affymax "VLSIPS" method) been applied. The second method, called coded mixture synthesis is known, uses nucleotides, peptides or other types of more inert chemical markers to identify each compound.

During one Deconvolution, the third approach, is a series of compound blends combinatorially synthesized, with some specific ones each time Structural features are fixed. Each mixture is considered a mixture tested and tracked the most active combination. Fix further rounds other structural features systematically, up to a manageable number of discrete Structures can be synthesized and screened. Researchers who working with peptides, for example, can use deconvolution, the most active peptide sequence from millions of possibilities to optimize or locate.

In a related field, surfaces that have been modified to provide reactive groups or other desired functionality have long been used to carry out solid-phase syntheses of both polymeric and non-polymeric molecules. A variety of feast Phase resins are commercially available, eg, those available from Argonaut Technologies, including their product line of polystyrene, ArgoGel ^™, and ArgoPore ^™ resins. Along with similar product lines, published International Patent Application WO9727226 ("Highly Functionalized Polyethylene Glycol Grafted Polystyrene Supports"), assigned to Argonaut Technologies, discloses polymers and graft copolymers having a backbone of poly (methylstyrene) and side chain polymers of poly (ethylene oxide).

Such carrier are also reported in JW Labadie ("Polymeric Supports for Solid Phase Synthesis "), Current Opinions in Chemical Biology 2: 346 (1998). This For example, article describes the way in which functional Groups can be introduced in slightly crosslinked polystyrene, under Use of either functional styrenic monomers or in one Nachfunktionalisierungsschritt. In both approaches, however, are the functional Groups apparently attached to the polystyrene polymers used be to the wearer self (e.g., sphere), e.g. contrary to that they as a separate coating to a pre-existing carrier added become.

The article by Labadie also describes the use of PEG graft polystyrene, eg in the form of the "TentaGel" product made by grafting ethylene oxide onto hydroxyl functional polystyrene. "The article also describes the manner in which various" Disadvantages associated with PEG graft resins have been overcome by a graft resin identified as "ArgoGel ^™ " which is characterized as branching at the polystyrene graft linkage by the use of a polystyrene diol as the base resin According to these approaches, the resulting polymers appear to be limited to functional groups at their terminal ends, as opposed to having functional groups in multiple positions along the length of the polymer.

On Yet another area is a variety of polymeric compositions for use as electrophoretic gels. Please refer generally Righetti et al., J. Chromatog. B. Biomed. Sci. 10; 699 (1-2): 63-75 (1997), which describes recent advances in polyacrylamide gel electrophoresis.

Please refer e.g. 5,470,916 for "Formulations for Polyacrylamide Matrices in Electrokinetic and Chromatographic Methodologies ". U.S. No. 5,470,916 describes preparations which are known via polymerization or Copolymerization of a unique class of monomers obtained become.

Please refer also U.S. 5,785,832 for "Covalently Cross-linked, mixed-bed agarose polyacrylamide matrices for electrophoresis and Chromatography ", which describes polyacrylamide matrices based on a new class of N-mono and di-substituted acrylamide monomers. U.S. 5,785,832 describes the manner in which mixed bed matrices of the polyacrylamide-agarose type, covalently linked (crosslinked) in the separation of fragments of nucleic acid, certain DNA, an intermediate size (of 50 to 5,000 base pairs) and of high molecular weight proteins Mass (> 500,000 Da) are suitable. U.S. 5,785,832 provides covalently linked mixed bed matrices of polyacrylamide agarose suitable for use in the separation of Fragments of nucleic acids a medium size suitable are.

substituted Polyacrylamides such as those described above are for use in the preparation of electrophoretic gels limited and, as Applicant knows, have not yet been attached to surfaces, let alone attached been for the purposes of providing a solid phase synthesis surface, or by photochemical means.

On a separate area has the legal successor of the present The invention has previously been the modification of surfaces for a variety of purposes described and using a variety of reagents. In particular, close these reagents generally involve the use of photochemistry, and especially photoreactive groups, e.g. for the addition of polymers and others molecules on support surfaces. Please refer e.g. 4,722,906, 4,979,959, 5,217,492, 5,512,329, 5,563,056, 5,637,460, 5,714,360, 5,741,551, 5,744,515, 5,783,502, 5,858,653 and 5,942,555.

SUMMARY THE INVENTION

• a) Bereitstellen eines Trägermaterials mit einer Oberfläche, die für den Einsatz in kombinatorischen Synthesen eingerichtet ist,
• b) Bereitstellen eines polymeren Reagenzes, gebildet durch die Polymerisation von Monomeren der Formel:
  
  wobei R₁ Wasserstoff oder C₁-C₆-Alkyle bedeutet, und wobei R₂ und R₃, unabhängig voneinander, Wasserstoff, C₁-C₆-Alkyle oder Phenyle enthaltend einen oder mehrere reaktive Substituenten ausgewählt aus
  
  OR₅ oder SR₅ bedeuten (wobei R₄ ein C₁-C₆-Alkyl oder ein heterocyclischer Ring enthaltend ein oder mehrere Stickstoffatome ist, und R₅ ein C₁-C₆-Alkyl oder Phenyl ist, enthaltend einen oder mehrere reaktive Substituenten ausgewählt aus
  
• c) Applizieren des Reagenzes auf die Trägeroberfläche und kovalentes Anbinden des polymeren Reagenzes an die Trägeroberfläche,
• d) Bereitstellen eines ersten reaktiven Monomers, das zur Festphasensynthese eingerichtet ist, z.B. ausgewählt aus Nukleotiden und Aminosäuren, wobei das Monomer eine korrespondierende Gruppe umfaßt, die thermochemisch mit dem gebundenen reaktiven Substituenten reaktiv ist, und ebenfalls bevorzugt eine oder mehrere Gruppen umfaßt, die mit einer nachfolgenden zweiten Monomereinheit im Verlauf der Festphasensynthese reaktiv ist,
• e) Kontaktieren und Reagieren des ersten Monomers mit dem polymeren Reagenz auf der Trägeroberfläche unter Bedingungen, die geeignet sind zum Umsetzen der korrespondierenden Gruppe mit dem gebundenen reaktiven Substituenten, um so eine wachsende polymere Kette bereitzustellen, und
• f) Bereitstellen und nachfolgendes Anbringen weiterer Monomere an die wachsende Polymerkette, um ein gewünschtes Polymerprodukt zu erhalten.

The present invention provides a method of performing a solid phase synthesis, the method of claim 1 comprising the steps of:

• a) providing a support material having a surface adapted for use in combinatorial syntheses,
• b) providing a polymeric reagent formed by the polymerization of monomers of the For mel:
  
  wherein R _{1 is} hydrogen or C ₁ -C ₆ alkyls, and wherein R ₂ and R ₃ , independently of one another, are hydrogen, C ₁ -C ₆ -alkyls or phenyls containing one or more reactive substituents selected from
  
  OR ₅ or SR ₅ (wherein R _{4 is} a C ₁ -C ₆ alkyl or a heterocyclic ring containing one or more nitrogen atoms, and R _{5 is} a C ₁ -C ₆ alkyl or phenyl containing one or more reactive substituents selected from
  
• c) applying the reagent to the carrier surface and covalently bonding the polymeric reagent to the carrier surface,
• d) providing a first reactive monomer adapted for solid phase synthesis, eg selected from nucleotides and amino acids, the monomer comprising a corresponding group which is thermochemically reactive with the attached reactive substituent, and also preferably comprising one or more groups substituted with a subsequent second monomer unit is reactive in the course of the solid phase synthesis,
• e) contacting and reacting the first monomer with the polymeric reagent on the support surface under conditions suitable for reacting the corresponding group with the attached reactive substituent so as to provide a growing polymeric chain, and
• f) providing and subsequently attaching further monomers to the growing polymer chain to obtain a desired polymer product.

As soon as it is formed in this way, the resulting polymeric Product either withheld and used in situ (e.g., in its bound state), or it can be cut from its position on the carrier and removed be used in a different way.

In In another aspect, the present invention provides a polymeric reagent composition adapted to be supported on a support surface to become this surface to provide a high density of reactive groups. The way coated surface can for Any suitable purpose will be used and is especially good suitable for use as a solid phase synthesis support surface. The Synthetic carrier surface can again in repetitive and combinatorial syntheses, such as Synthesis of polynucleotides, polypeptides and polysaccharides, be used. The polymeric coating can be used an enlarged effective surface especially in situations where the effective area on the surface itself is limited, as on the surface of a sphere or a sphere Silicon wafer. In this way, the polymeric coating provides an optimal combination of such properties as the density reactive groups and the surface or the volume.

In a preferred embodiment For example, the polymeric reagent will be in the form of a hydrophilic or amphiphilic polymeric reagent adapted to act on a support surface via stable covalent Bindings become coated to the surface with a high, however To provide controllable density of functional groups, the Solid phase synthesis of peptides, oligonucleotides and other oligomers (e.g., peptide nucleic acids) or non-polymeric organic compounds.

wobei R₁ Wasserstoff oder C₁-C₆-Alkyle darstellt und wobei R₂ und R₃, unabhängig voneinander, Wasserstoff, C₁-C₆-Alkyle oder Phenyle darstellen, die eine oder mehrere reaktive Substituenten enthalten, die ausgewählt sind aus

OR₅ oder SR₅ (wobei R₄ ein C₁-C₆-Alkyl oder ein heterocyclischer Ring enthaltend ein oder mehrere Stickstoffatome ist, und R₅ ein C₁-C₆-Alkyl oder Phenyl ist, enthaltend einen oder mehrere reaktive Substituenten ausgewählt aus

The reagent can be prepared by the polymerization of one or more monomers containing functional groups of the formula:

wherein R _{1 is} hydrogen or C ₁ -C ₆ alkyls and wherein R ₂ and R ₃ , independently of one another, are hydrogen, C ₁ -C ₆ alkyls or phenyls containing one or more reactive substituents selected from

OR ₅ or SR ₅ (wherein R _{4 is} a C ₁ -C ₆ alkyl or a heterocyclic ring containing one or more nitrogen atoms, and R _{5 is} a C ₁ -C ₆ alkyl or phenyl containing one or more reactive substituents selected out

Certain monomers of this type are described, for example, in Righetti ('832), the disclosure of which is incorporated herein by reference. In a preferred embodiment of the present invention, the polymeric reagent is prepared from monomers including N-acryloyl-amino-ethoxy-ethanol (AAEE), a highly hydrophilic monomer that is highly resistant to hydrolysis (Chiari, Micheletti, Nesi, Fazio , Righetti, Electrophoresis 15, 1994, 177-186). Other monomers of this type are disclosed in U.S. Pat US 5,858,653 described, the disclosure of which is incorporated herein by reference.

The Method and reagent of this invention finds particular application in situations where it is desired is, the synthesis capacity to increase, without necessarily a corresponding or excessive increase of the reaction volume needed becomes. The reagent of the present invention provides a preformed Polymer composition in which the polymer molecules purified, characterized and can be controlled in a way that was previously not possible is.

In a preferred embodiment includes the reagent is the addition of preformed synthetic polymers on a surface (what distinguishes it from those generated by in situ polymerization the carrier are formed), in particular the addition of the preformed polymers by photochemical means. It is also preferred that the functional Groups at a variety of positions along the polymer backbone available. The number (or average number) and the position The functional groups can be determined by the choice of monomers used be controlled to form the polymer, e.g. by the ratio of functional group-containing monomers to "diluent" monomers.

A Polymer reagent composition of this invention provides optimum Combination of such properties as swellability, density functional Groups, reactivity, Permeability, Hydrophilicity and hydrolytic stability. In a particularly preferred embodiment comprises the reagent composition providing a polymeric derivative one or more different reactive groups. The reagent composition can to the surface be attached in any suitable manner, and it is preferred covalently on the surface tethered, more preferably by the use of photoreactive Groups.

suitable support materials shut down Balls, foils, wafers, films, discs and plates (e.g., microbore plates) prepared from such materials as organosilane-treated glass, Organosilane-treated silicon, polypropylene, polyethylene and Polystyrene (optionally cross-linked with divinylbenzene). additional support materials shut down grafted polyacrylamide beads, latex beads, dimethylacrylamide beads (optionally crosslinked with N, N'-bis-acryloylethylenediamine), Glass particles coated with hydrophobic polymers, etc. (i.e. with a rigid or semi-rigid surface). divinylbenzene-crosslinked, Polyethylene glycol grafted polystyrene-like beads may also be used.

In a particularly preferred embodiment, the reagent comprises a hydroxyl-substituted polyacrylamide reagent. Such a reagent may be present on a surface, for example, in a photochemical manner desired manner and concentration are added to provide the surface with a desired density of reactive groups (eg primary hydroxyl groups).

One Polymer of this invention can be prepared using any suitable means, e.g. by the reaction of monomers, which provide one or more functional groups, with one or more reactive comonomers (e.g., monomers which are a photoreactive Provide group) and / or with one or more non-reactive Comonomers (e.g., "diluent" monomers that are not photoreactive) Group or functional group). Professionals in the field are based on the present description the way to which a polymer of this invention is free radical polymerized using concentrations and ratios of monomers, tailored, to the desired surface properties can be synthesized. Thus, the relative and absolute Concentrations of functional groups and the molecular weight of the polymer (and the extent of Branching, etc.) and the means for immobilizing the polymer (Such as the number and / or locations of the photoactivatable groups along its length) all set to optimize performance.

comonomers with functional groups of different types and reactivities can also selected become. Although it is not the only determining factor, it can the length any spacer that is between a functional group and the eventual Polymer chain is included, a predictable or determinable effect on the reactivity of the functional group to have. In addition, relatively inert Monomers are included to act as diluent monomers to to adjust the density of the functional groups to desired levels and the desired ones To achieve polymer properties, e.g. its hydrophilic, hydrophobic or amphiphilic nature, which in turn determines its dissolution properties can influence.

Finally, comonomers Also included are the reactive groups for immobilization of the polymer on a surface provide. Such monomers preferably contain photoactivatable Groups or can include thermochemically reactive groups that can be used to either the polymer directly at a corresponding reactive site or group on the surface to add, or to another reagent which itself is a photoactivatable group provides. For example, you can.

hydroxyl activated with a variety of activating agents (e.g. 1,1-carbonyldiimidazole 2,2,2-trifluoroethanesulfonyl chloride or 2-fluoro-1-methylpyridinium p-toluenesulfonate). Such reactions can used to contain a hydroxyl polymer on a surface To immobilize amine groups (e.g., glass-coated with 3-aminopropyltriethoxysilane). In this example, any excess can be activated Hydroxyl groups that do not react with amines on the surface, back be hydrolyzed to free hydroxyl groups. The comonomers can also be selected at different polymerization rates, to optimize the distribution of comonomers in the polymer. optional or additionally can the comonomer distribution through the manufacture and use be controlled and influenced by block copolymers.

Hydrophilic or amphiphilic polymers also be provided with immobilization agents on a surface over stable covalent bonds and multiple functional groups of the type described herein Type. Such polymers find particular application for solid phase synthesis of peptides, oligonucleotides, similar Types of polymers (e.g., peptide nucleic acids) and non-polymeric organic ones Links. The use of presynthesized polymers, e.g. in contrast to grafted polymers or those that are in situ provide a number of advantages including Ability, to purify and characterize the polymer prior to immobilization.

One A method for providing reactive groups on a surface is also disclosed. The process includes the step of coating the surface with a reagent composition described herein.

DETAILED DESCRIPTION

One Reagent of this invention can be prepared by the polymerization of monomers containing functional groups, optional and preferred in combination with other monomers, such as those others Groups, diluent monomers and the like.

wobei R₁ Wasserstoff oder C₁-C₆-Alkyle darstellt und wobei R₂ und R₃, unabhängig voneinander, Wasserstoff, C₁-C₆-Alkyle oder Phenyle darstellen, die einen oder mehrere reaktive Substituenten enthalten, die ausgewählt sind aus

OR₅ oder SR₅ (wobei R₄ ein C₁-C₆-Alkyl oder ein heterocyclischer Ring enthaltend ein oder mehrere Stickstoffatome ist, und R₅ ein C₁-C₆-Alkyl oder Phenyl ist, enthaltend einen oder mehrere reaktive Substituenten ausgewählt aus

Optional und bevorzugt werden die resultierenden Polymere ebenfalls an der Oberfläche über photochemische Mittel, z.B. durch die Integration einer oder mehrerer Photogruppen in das Polymer über Photogruppen-enthaltende copolymerisierbare Monomere angefügt.A polymer of the present invention can be prepared by polymerizing one or more monomers selected from the group:

wherein R _{1 is} hydrogen or C ₁ -C ₆ -alkyls and wherein R ₂ and R ₃ , independently of one another, are hydrogen, C ₁ -C ₆ -alkyls or phenyls which contain one or more reactive substituents which are selected from

OR ₅ or SR ₅ (wherein R _{4 is} a C ₁ -C ₆ alkyl or a heterocyclic ring containing one or more nitrogen atoms, and R _{5 is} a C ₁ -C ₆ alkyl or phenyl containing one or more reactive substituents selected out

Optionally and preferably, the resulting polymers are also added to the surface via photochemical means, for example by the integration of one or more photo-groups in the polymer via photogroup-containing copolymerizable monomers.

comonomers can selected be to any desired To provide property or function, including any desired Reactivity. Although it is not the only determining factor, the length of the Spacer between the functional groups and the polymer main chain often a Influence on the reactivity of functional groups.

In addition, "inert" or "dilution" monomers may be used be to the density of functional groups at optimal levels set and desired To obtain polymer properties, such as hydrophilic or amphiphilic Polymers for optimal solvation properties. Examples include such monomers for example acrylamide, N-vinylpyrrolidone, methacrylamide, N-isopropylacrylamide, N-vinylpyridine, N-vinyl caprolactam, Styrene, vinyl acetate and N-acryloylmorpholine.

A preferred composition of this invention includes one or more pendant, latently reactive (preferably photoreactive) groups that are covalently attached or adapted to be attached, directly or indirectly, to a copolymerizable monomer. Photoreactive groups are defined herein and preferred groups are sufficiently stable to be stored under conditions in which they retain such properties. See for example US 5,002,582 the disclosure of which is incorporated herein by reference. Latent reactive groups may be selected that are sensitive to different regions of the electromagnetic spectrum, with those that are sensitive to ultraviolet and visible regions of the spectrum (referred to herein as "photoreactive").

Photo Reactive Groups respond to specific external stimuli applied to active species production with resulting covalent bonding to be subject to an adjacent chemical structure, as for example provided by the same or a different molecule. Photo Reactive Groups are such groups of atoms in a molecule that their keep covalent bonds unchanged under storage conditions, however, covalent upon activation by an external source of energy Bonds with other molecules form.

The Photoreactive groups produce active species, such as free radicals and in particular nitrenes, carbenes and excited states of Ketones upon absorption of electromagnetic energy. Photo Reactive Groups can selected to address different areas of the electromagnetic spectrum, and photoreactive groups, for example, on ultraviolet and sensitive regions of the spectrum are preferred and can sometimes referred to herein as "photochemical Group "or" photogroup ".

Photo Reactive Aryl ketones are preferred, such as acetophenone, benzophenone, anthraquinone, Anthrone and anthrone-like heterocycles (i.e., heterocyclic analogs by Anthron, such as those with N, O or S in the 10-position), or their substituted (e.g., ring-substituted) derivatives. Examples preferred aryl ketones include heterocyclic derivatives by Anthron, including Acridone, xanthone and thioxanthone, and their ring-substituted Derivatives.

The functional groups of such ketones are preferred because they are light capable of undergoing an activation / deactivation / reactivation cycle described herein to be subject. Benzophenone is a particularly preferred photoreactive Unity, as it is capable of photochemical excitation the initial one Formation of an excited singlet state, that of an intersystem transition is subject to the triplet state. The excited triplet state may be in carbon-hydrogen bonds by abstraction of a hydrogen atom insert (for example, from a support surface), whereby a radical pair is generated. A subsequent collapse of the radical pair to form a new carbon-carbon bond. When a reactive bonding (e.g., carbon-hydrogen) to the bond is not available is the ultraviolet light induced excitation of the Benzophenone group reversible and the molecule returns to its ground state energy level when removing the power source back. Photoactivatable aryl ketones, like benzophenone and acetophenone, are of particular importance since these groups are subject to multiple reactivation in water are and therefore an increased Provide coating efficiency.

The azides form an additional preferred class of photoreactive groups and include aryl azides (C ₆ R ₅ N ₃ ) such as phenyl azide and especially 4-fluoro-3-nitrophenyl azide, acyl azides (-CO-N ₃ ) such as benzoyl azide and p-methylbenzoyl azide, Azido formates (-O-CO-N ₃ ) such as ethyl azidoformate, phenyl azidoformate, sulfonyl azides (-SO ₂ -N ₃ ) such as benzenesulfonyl azide, and phosphoryl azides (RO) ₂ PON ₃ such as diphenylphosphoryl azide and diethylphosphoryl azide. Diazo compounds form another class of photoreactive groups and include diazoalkanes (-CHN ₂ ) such as diazomethane and diphenyldiazomethane, diazoketones (-CO-CHN ₂ ) such as diazoazetophenone and 1-trifluoromethyl-1-diazo-2-pentanone, diazoacetates (-O -CO-CHN ₂ ) such as t-butyldiazoacetate and phenyldiazoacetate, and beta-keto-alpha-diazoacetate (-CO-CN ₂ -CO-O-) such as t-butyl-alpha-diazetoacetate. Other photoreactive groups include the diazirines _(-CHN2) such as 3-trifluoromethyl-3-phenyldiazirine, and ketenes (-CH = C = O) such as ketene and diphenylketene.

at Activation of the photoreactive groups makes the reagent molecules covalent to each other and / or to the material surface through covalent bonds bound by residues of the photoreactive groups. Exemplary photoreactive Groups and their residues upon activation are shown as follows (where R and R 'any non-interfering organic groups are).

The Photoactivatable monomers of the invention may be applied to any surface Carbon-hydrogen bonds are applied with which the photoreactive groups can react to the resulting Polyacrylamide on surfaces to immobilize. Examples of suitable substrates include but are not limited to polypropylene, polystyrene, poly (vinyl chloride), Polycarbonate, poly (methyl methacrylate), parylene and any of numerous organosilanes that are used to glass or others organic surfaces pretreat.

polymers of this invention are preferred by free radical polymerization synthesized using concentrations and ratios of monomers, the tailor made are the ones you want surface properties to achieve. Thus, you can the contents of functional groups, the molecular weight of the polymer and the means for immobilizing the polymer (e.g., by integration of photoactivatable groups) by those skilled in the art become any desired product to obtain and / or the performance or the physical / chemical To optimize properties in one or more relationships.

One Reagent of the present invention can be used in a variety of ways be used to functionalized carrier surfaces for use in the To provide solid phase synthesis. In one embodiment the reagent may be packaged and provided separately, and, optionally, in bulk to be placed on a surface by the user To be applied at the time of use. In another embodiment The reagent can be applied and covalently bound to a support (e.g., by photochemical means) at the time of manufacture of the carrier itself, and the resulting coated substrate can be packaged and sold in a form that is essentially for use is done.

TABLE OF STRUCTURES

VERBINDUNG I

CONNECTION I

VERBINDUNG II

CONNECTION II

VERBINDUNG III

CONNECTION III

VERBINDUNG IV

CONNECTION IV

VERBINDUNG V

CONNECTION V

VERBINDUNG VI

CONNECTION VI

VERBINDUNG VII

CONNECTION VII

VERBINDUNG VIII

CONNECTION VIII

• where x = 0 to 5 mol%, y = 5 to 100 mol% and z = 0 to 95 mol%

VERBINDUNG IX

CONNECTION IX

• where x = 0 to 5 mol%, y = 5 to 100 mol% and z = 0 to 95 mol%

VERBINDUNG X

CONNECTION X

• where m = 15 to 45 and n = 50 to 150 (occurring statistically)

VERBINDUNG XI

CONNECTION XI

• where x = 0 to 5 mol%, y = 5 to 100 mol% and z = 0 to 95 mol%

The The following examples are provided to illustrate the present invention to illustrate, but not to limit. While the Invention has been described with respect to their preferred embodiments It will be appreciated by those skilled in the art that the invention with modifications made within the spirit and scope of the appended claims can.

EXAMPLES

example 1

Preparation of 4-benzoylbenzoyl chloride (BBA-Cl) (compound I)

To prepare a reactive photogroup, 4-benzoylbenzoic acid (BBA), 1.0 kg (4.42 mol), was added to a dry 5 liter Morton flask equipped with reflux condenser and overhead stirrer, followed by the addition of 645 ml (8 , 84 mol) of thionyl chloride and 725 ml of toluene. Dimethylformamide (3.5 ml) was then added and the mixture heated at reflux for 4 hours. After cooling, the solvents were removed under reduced pressure and the remaining thionyl chloride by three evapor removed using 3 × 500 ml of toluene. The product was recrystallized from 1: 4 toluene: hexane to give 988 g (91% yield) after drying in a vacuum oven. The product melting point was 92-94 ° C. Nuclear Magnetic Resonance Analysis (NMR) at 80 MHz ( ¹ H NMR (CDCl ₃ )) was consistent with the desired product. Aromatic protons 7,20-8,25 (m, 9H). All chemical shift values are in ppm downfield from an internal tetramethylsilane standard. The final compound was stored for use in the preparation of a monomer for use in the synthesis of photoactivatable polymers, for example as described in Example 3.

Example 2

Preparation of N- (3-amino-propyl) -methacrylamide hydrochloride (AMPA) (compound II)

To form an amine-containing monomer intermediate, a solution of 1,3-diaminopropane, 1,910 g (25.77 moles) in 1,000 mL of CH ₂ Cl _{2, was added} to a 12 liter Morton flask and cooled in an ice bath. A solution of t-butylphenyl carbonate, 1,000 (5.15 mol), in 250 mL of CH ₂ Cl ₂ was then added dropwise at a rate keeping the reaction temperature below 15 ° C. Following the addition, the mixture was warmed to room temperature and stirred for 2 hours. The reaction mixture was diluted with 900 mL of CH ₂ Cl ₂ and 500 g of ice, followed by the slow addition of 2,500 mL of 2.2 N NaOH. After testing to ensure that the solution was basic, the product was transferred to a separatory funnel and the organic layer removed and set aside as Extract # 1. The aqueous solution was then extracted with 3 x 1250 mL of CH ₂ Cl ₂ , each extraction being saved as a separate fraction. The four organic extracts were then washed successively with a single amount of 1250 ml of 0.6N NaOH starting with fraction # 1 and proceeding to fraction # 4. This washing was repeated a second time with a fresh amount of 1250 ml of 0.6 N NaOH. The organic extracts were then combined and dried over Na ₂ SO ₂ . Filtration and evaporation of the solvent to a constant weight gave 825 g of N-mono-t-BOC-1,3-diaminopropane, which was used without further purification.

A solution of methacrylic anhydride, 806 g (5.23 mol) in 1020 ml CHCl ₃ , was placed in a 12 liter Morton flask equipped with overhead stirrer and cooled in an ice bath. Phenothiazine, 60 mg, was added as an inhibitor, followed by the dropwise addition of N-mono-t-BOC-1,3-diaminopropane, 825 g (4.73 mol) in 825 mL of CHCl ₃ . The rate of addition was controlled to always keep the reaction temperature below 10 ° C. After the addition was complete, the ice bath was removed and the mixture allowed to stir overnight. The product was diluted with 2400 ml of water and transferred to a separatory funnel. After thorough mixing, the aqueous layer was removed and the organic layer was washed with 2400 ml of 2N NaOH, which ensured that the aqueous layer was basic. The organic layer was then dried over Na ₂ SO ₄ and filtered to remove the drying agent. A portion of the CHCl ₃ solvent was removed under reduced pressure until the combined weight of the product and the solvent was about 3,000 g. The desired product was then precipitated by slow addition of 11.0 L of hexane to the stirred CHCl ₃ solution, followed by overnight storage at 4 ° C. The product was isolated by filtration and the solid was rinsed twice with a solvent combination of 900 mL of hexane and 150 mL of CHCl ₃ . Thorough drying of the solid gave 900 g of N- [N '- (t-butyloxycarbonyl) -3-aminopropyl] methacrylamide, m.p. 85.8 ° C, by differential scanning calorimetry (DSC). An analysis on an NMR spectrometer was consistent with the desired product: ¹ H-NMR _(CDCl3) amide NH's 6.30 to 6.80, 4.55 to 5.10 (m, 2H), vinyl protons 5.65 , 5.20 (m, 2H), methylene adjacent to N 2.90-3.45 (m, 4H), methyl 1.95 (m, 3H), residual methylene 1.50-1.90 (m, 2H ) and t-butyl 1,40 (s, 9H).

A 2-liter, 3-neck round bottom flask was equipped with an overhead stirrer and a gas sparger. Methanol, 700 ml, was added to the flask and cooled on an ice-bath. While stirring, HCl gas was bubbled into the solvent at a rate of about 5 L / min for a total of 40 minutes. The molarity of the final HCl / MeOH solution was determined to be 8.5 M by titration with 1 N NaOH using phenolphthalein as an indicator. The N- [N '- (t-butyloxycarbonyl) -3-aminopropyl] methacrylamide, 900 g (3.71 mol), was added to a 5 liter Morton flask equipped with an overhead stirrer and a gas outlet adapter, followed by the addition of 1150 ml of methanol solvent. Some solids remained in the flask with this volume of solvent. Phenothiazine, 30 mg, was added as an inhibitor, followed by the addition of 655 ml (5.57 mol) of the 8.5 M HCl / MeOH solution. The solids slowly dissolved with the evolution of gas, but the reaction was not exothermic. The mixture was stirred overnight at room temperature to ensure a complete reaction. Any solids were then removed by filtration and an additional 30 ml of phenothiazine added. The solvent was then stripped under reduced pressure and the resulting solid residue washed 3 times 1000 ml of isopropanol azeotroped with evaporation under reduced pressure. Finally, the product was dissolved in 2,000 ml of isopropanol under reflux and 4,000 ml of ethyl acetate was added slowly with stirring. The mixture was allowed to cool slowly to room temperature and was stored at 4 ° C overnight. Compound II was isolated by filtration and dried to constant weight, yielding 630 g with a melting point of 124.7 ° C by DSC. Analysis on an NMR spectrometer was consistent with the desired product: ¹ H NMR (D ₂ O) vinyl protons 5.60, 5.30 (m, 2H), methylene adjacent to amide N 3.30 (t, 2H ), Methylene adjacent to amine N 2.95 (t, 2H), methyl 1.90 (m, 3H) and residual methylene 1.65-2.10 (m, 2H). The final product was stored for use in the preparation of a monomer used in the synthesis of photoactivatable polymers, such as described in Example 3.

Example 3

Preparation of N- [3- (4-benzoylbenzamido) propyl] methacrylamide (BBA-APMA) (compound III)

The reactive photogroup of Example 1 and the amine monomer of Example 2 were reacted (via an amide linkage) to form a photogroup-containing monomer in the following manner. Compound II 120 g (0.672 mol), prepared according to the general method described in Example 2, was added to a dry 2-liter, 3-neck round bottom flask equipped with an overhead stirrer. Phenothiazine, 23-25 mg, was added as an inhibitor, followed by 800 ml of chloroform. The suspension was cooled to below 10 ° C on an ice bath and 172.5 g (0.705 mol) of Compound I, prepared according to the general method described in Example 1, was added as a solid. Triethylamine, 207 ml (1.485 mol) in 50 ml of chloroform was then added dropwise over a period of 1-1.5 hours. The ice bath was removed and stirring continued at ambient temperature for 2.5 hours. The product was then washed with 600 ml of 0.3 N HCl and 2 x 300 ml of 0.07 N HCl. After drying over sodium sulfate, the chloroform was removed under reduced pressure and the product was recrystallized twice from 4: 1 toluene: chloroform using 23-25 mg of phenothiazine in each recrystallization to prevent polymerization. Typical yields of compound III were 90% with a melting point of 147-151 ° C. Analysis on an NMR spectrometer was consistent with the desired product: ¹ H-NMR (CDCl ₃ ) aromatic protons 7.20-7.95 (m, 9H), amide NH 6.55 (broad t, 1H), Vinyl protons 5.65, 5.25 (m, 2H), methylene adjacent to amide N's 3.20-3.60 (m, 4H), methyl 1.95 (s, 3H), and residual methylene 1.50-2 , 00 (m, 2H). The final product was stored for use in the synthesis of photoactivatable polymers, as described, for example, in Examples 8-11 and 13.

Example 4

Preparation of N- (3-hydroxypropyl) acrylamide (HPA) (Compound IV)

One Monomer containing a functional group in the form of a hydroxyl group was made in the following way. acryloyl, 53 ml (0.66 mol) was cooled to -40 ° C in a Three-necked round bottom flask immersed in an isopropanol / dry ice bath, cooled. The flask was equipped with an addition funnel, a thermometer and equipped with an argon inlet. ethanol (1000 ml) was cooled to -40 ° C and added to the cold acryloyl chloride. 3-amino-1-propanol, 100 ml (1.3 mol) was dissolved in 1000 ml of ethanol. This solution was transferred to the addition funnel and added dropwise to the acryloyl chloride. The solution was in the ethanol / dry ice bath for Stirred for 2 hours, followed by a stir at 4 ° C overnight. The solvent was evaporated. After the addition of 25 ml of phenothiazine, the Residue dissolved in chloroform and purified on silica gel. After washing with chloroform was the product eluted with acetone to give 96.1 g (82% yield).

Example 5

Preparation of N- (2-ethoxy- (2-hydroxyethyl) acrylamide (HEEA) (connection V)

One Another monomer containing a hydroxyl group was replaced by the essentially the same procedure as in Example 4, except that 40.4 ml (0.5 mol) of acryloyl chloride with 100.2 ml (1.0 mol) of 2- (2-aminoethoxy) ethanol have been implemented. The compound was silica gel with acetone eluted to give 94.6 g (119% yield).

Example 6

Preparation of N-acrylamido-D-glucosamine (AGA) (Compound VI)

One another alternative monomer containing a functional group in the form of a hydroxyl group was in the following way and Made way. Glucosamine hydrochloride, 10.0 g (0.0464 mol), was added to 12 ml of 3.8 M sodium hydroxide. Potassium carbonate. 0.30 g (0.0022 mol) and sodium nitrite, 0.35 g (0.0051 mol) then added and the mixture stirred until a clear solution was obtained has been. To the clear solution 10 ml of chloroform were added and the mixture became vigorous touched, while she was in an ice bath. A solution of acryloyl chloride, 4.45 g (4.0 ml, 0.0492 mol) in 5 ml of chloroform was in proportions of 100 microliters with alternate additions of 55 μl Add 10N sodium hydroxide (4.95 ml total) with stirring in an ice bath. Stirring was for Continued for 30 minutes, after the additions were completed. The chloroform was removed. The pH was adjusted to 3, followed by 3 x 10 ml Extracts of chloroform to remove acrylic acid. The water solution became at about Stored at 4 ° C. The use of AGA in the polymerization is illustrated in Examples 12 and 13 described.

Example 7

Preparation of N- (2-mercaptoethyl) -2,6-bis (4-benzoylbenzamido) hexanamide (Compound VII)

A photoactivatable chain transfer reagent was prepared in the following manner. Lysine monohydrochloride, 3.65 g (20 mmol), was dissolved in 8 mL of 2N sodium hydroxide and cooled in an ice bath. A solution of 10.77 g (44 mmol) of 4-benzoylbenzoyl chloride prepared according to the procedure generally described in Example 1 in 17 ml of chloroform was added simultaneously with 4.48 g of sodium hydroxide in 19 ml of water. The reaction was stirred on the ice bath for 2 hours and then allowed to warm to room temperature for 3 hours. Hydrochloric acid was used to adjust the pH to 1 and an additional 60 ml of chloroform was added. A centrifuge was used to separate the layers, and the aqueous layer was extracted with 3 x 50 ml of chloroform. The combined organic extracts were dried over sodium sulfate. An approach was carried out to recrystallize the resulting solid product from 80% acetic acid, but the recovery of the product was poor. The mother liquors were diluted with water to precipitate the product which was then dissolved in chloroform and washed with 10% sodium bicarbonate, 1N hydrochloric acid and finally water. The solution was dried over sodium sulfate and the product used without purification. Analysis on an NMR spectrometer was consistent with the desired product: ¹ H-NMR _(CDCl3) acid proton 8.45 (broad s, 1H), aromatic and amide protons 7.00 to 8.10 (m, 20H) , CH 4.50-4.90 (m, 1H), methylene adjacent to N 3.30-3.70 (m, 2H), remaining methylenes 1.10-2.25 (m, 6H).

The Lysine derivative, 4.35 g (7.73 mmol), and N-hydroxysuccinimide, 0.901 g (7.83 mmol) were dissolved in 40 ml of dry 1,4-dioxane, followed from the addition of 1.951 g (9.45 mmol) of 1,3-dicyclohexylcarbodiimide (DCC) in 10 ml of 1,4-dioxane. The mixture was allowed to stand overnight at room temperature touched become. The resulting white Solid was filtered off and washed with 2 x 25 ml of 1,4-dioxane. The solvent was removed under reduced pressure and the residue with 3 × 25 rinsed in hexane, to excess DCC too remove. The resulting N-oxysuccinimide ester (NOS), 4.10 g (81% yield) was used without further purification.

2-Aminoethanethiol hydrochloride, 0.75 g (6.6 mmol), was diluted with 15 ml of chloroform and 1.09 ml of triethylamine under argon atmosphere. The NOS ester, 4.10 g (6.22 mol) in 25 ml of chloroform was added dropwise at room temperature over a period of 30 minutes. After 4 hours of reaction, the mixture was washed with water and 0.05 N hydrochloric acid, followed by drying over sodium sulfate. The product was purified using silica gel flash chromatography using a solvent system of 95: 5 CHCl ₃ : CH ₃ OH to give 2.30 g of the product in a 59% yield. Analysis on an NMR spectrometer was consistent with the desired product: ¹ H-NMR (CDCl ₃ ) aromatic and amide protons 6.90-8.00 (m, 21H), CH 4.40-4.85 (m , 1H), methylene adjacent to N 3.00-3.75 (m, 4H), remaining methylenes 1.00-2.95 (m, 8H) and SH 1.40 (t, 1H).

Example 8

Preparation of copolymer of acrylamide BBA-APMA and (HPA) (random photo-PA-HPA) (Compound VIII)

One Photoactivatable copolymer of the present invention has been disclosed the following way. Acrylamide, 1.69 g (23.8 g mmol), for use as a "diluent" monomer, as is described herein in 43.5 ml of DMSO together with 0.17 g (0.49 mmol) BBA-AMPA (photogroup-containing monomer, Comp III), prepared according to the general in Example 3, 3.14 g (24.3 mmol) of HPA (a OH-containing monomer, compound IV) prepared according to the general in Example 4, 0.10 g (0.58 mmol) of 2,2'-azobisisobutyronitrile (AIBN) and 0.049 ml (0.32 mmol) of N, N, N ', N'-tetramethylethylenediamine (TEMED). The solution was using a helium gas tube for Deoxygenated for 4 minutes. The headspace was by argon replaced, and the vessel became for a Warm overnight at 55 ° C sealed. The reaction solution was in dialysis tubes (12-14000 MWCO) and opposite DI water for Dialyzed for 5 days. The water solution was lyophilized to give 4 g of white solid with a photo-loading of 0.094 μmol / mg (theory 0.097 μmol / mg) result.

Example 9

Preparation of a copolymer from BBA-APMA and (HPA) (statistical photo-HPA) (Compound VIII)

One Photoactivatable copolymer of the present invention has been disclosed the following manner (and without the use a "diluent" monomer such as acrylamide). BBA-AMPA (Compound III) prepared according to the procedure generally in Example 3, 0.17 g (0.49 mmol) was dissolved in 33 ml of DMSO along with 4.87 g (37.73 mmol) of HPA (Compound IV) according to the general in Example 4, 0.08 g (0.46 mmol) AIBN and 0.038 ml (0.25 mmol) TEMED dissolved. The solution was using a helium gas tube for Deoxygenated for 4 minutes. The headspace was replaced by argon, and the vessel became for a Stir over night at 55 ° C sealed. The reaction solution was in a dialysis tube (12-14000 MWCO) and dialyzed against DI water for 5 days. The water solution became lyophilized to 4.27 g white Solid with a photo-loading of 0.073 μmol / mg (theory 0.076 μmol / mg) result.

Example 10

Preparation of a copolymer from acrylamide BBA-APMA and (HEEA) (statistical photo-PA-HEEA) (Compound IX)

One The photoactivatable copolymer of the present invention has been disclosed in U.S. Pat made in the following manner. Acrylamide, 1.48 g (20.8 g mmol) was dissolved in 37.3 ml DMSO along with 0.15 g (0.42 mmol) BBA-AMPA (Compound III) prepared according to the general one in Example 3, 3.38 g (21.2 mmol) of HEEA (Compound V), manufactured according to the general in Example 5, 0.084 g (0.51 mmol) of AIBN and 0.043 ml (0.28 mmol) of TEMED. The solution was with a helium gas tube for 4 minutes removed from oxygen. The headspace was replaced by argon and the vessel for stirring overnight at 55 ° C sealed. The reaction solution was in a dialysis tube (12-14000 MWCO) and dialyzed against DI water for 5 days. The water solution became lyophilized to 3.7 g of white solid with a photo-loading of 0.092 μmol / mg (theory 0.085 μmolmg) too result.

Example 11

Preparation of a copolymer from BBA-APMA and (HEEA) (statistical photo-HEEA) (Compound IX)

One The photoactivatable copolymer of the present invention has been disclosed in U.S. Pat made in the following manner. BBA-APMA (Compound III), manufactured according to the general Example 3, 0.11 g (0.31 mmol) in 26 mL of DMSO along with 4.89 g (30.72 mmol) of HEEA (Compound V) prepared according to the general in Example 5, 0.06 g (0.37 mmol) AIBN and 0.031 ml (0.21 mmol) of TEMED was dissolved. The solution was using a helium gas tube for Deoxygenated for 4 minutes. The headspace was by argon replaced and the vessel for stirring overnight at 55 ° C sealed. The reaction solution was in a dialysis tube (12-14000 MWCO) and dialyzed against DI water for 6 days. The water solution became lyophilized to 4.35 g white To give solid, which as compound IX with a photo charge of 0.061 μmol / mg (Theory 0.062 μmol / mg) was designated.

Under Using the above procedure, a compound was similar to that described in the previous paragraph, however, crude HEEA was prepared instead of the purified HEEA used. The following ingredient loading was used to to give the Photo-HEEA (Compound IX). BBA-APMA (compound III), prepared according to the general Example 3, 0.043 g (0.12 mmol) in 10.4 mL of DMSO along with 1.96 g (12.29 mmol) of HEEA (Compound V) prepared according to the general in Example 5, 0.024 g (0.15 mmol) of AIBN and 0.12 ml (0.083 mmol) TEMED. The solution was with a helium gas tube for 4 minutes removed from oxygen. The headspace was replaced by argon and the vessel for stirring overnight sealed at 55 ° C. The reaction solution was in a dialysis tube (12-14000 MWCO) and dialyzed against DI water for 6 days. The water solution became lyophilized to 1.33 g white To give solid, which as compound IX with a photo charge of 0.078 μmol / mg (Theory 0.062 μmol / mg) was designated.

Example 12

Preparation of an end-point photoglucosamine polymer (Endpoint Di-BBA-AGA) (Compound X)

acrylamide (400 mg, 5.6 mmol) was dissolved in 10 ml of dimethyl sulfoxide (DMSO). To this solution 400 mg (1.48 mmol) of N-acryloylglucosamine (Compound VI) were added. additionally were 34 mg of N- (2-mercaptoethyl) -2,6-bis (4-benzoylbenzamido) hexanamide, 200 mg AIBN and 50 μl TEMED added. The solution was purged with nitrogen, then in a 55 ° C Oven over Arranged night. The resulting polymer solution was deionized Water using a SpectraPor I (Spectrum) dialysis membrane dialyzed. After dialysis, the solution was lyophilized. The obtained dried polymer was 0.58 wt. gm. At 0.1 mg / ml in deionized In water, the polymer had an absorbance at 265 nm of 0.202.

Example 13

Preparation of a copolymer from acrylamide, BBA-APMA and AGA (statistical photo-PA-AGA) (Compound XI)

To a solution of 200 mg (0.74 mmol) of N-acryloylglucosamine (compound VI) in 5 ml of DMSO, 500 mg acrylamide (7.0 mmol) 98 mg BBA-APMA (0.28 mmol, compound III), 50 mg azobiscyanovalerate and 100 μl TEMED. The solution was purged with nitrogen, then in a 55 ° C Oven over Arranged night. The polymer was dialyzed against deionized water and then lyophilized.

Example 14

Making a with PolyHPA-coated surface

One Polymer of PA is prepared using HPA monomer, synthesized according to example 4, and a polymerization reaction following the procedure from Example 9, except that the BBA-APMA was omitted. The lyophilized polymer is dissolved in a solution of 1,1'-carbonyldiimidazole in Formamide solved and could for react for an hour. glass slides are coated with aminopropyltrimethoxysilane, washed and dried. The activated polyHPA solution is modified to the amine Carrier applied and for incubated for one hour. The carriers are then dissolved in 0.1 M sodium carbonate solution containing 0.1 M ethanolamine for one Immersed to block remaining carbonylimidazole groups or to hydrolyze. The carriers are then washed in deionized water and dried.

Example 15

manufacturing a PhotopolyHEEA coated surface

A copolymer of HEEA (Compound V) and BBA-APMA (Compound III) prepared according to Example 10 is dissolved in deionized water at 2.0 mg / ml. Polystyrene microscope slides are immersed in the polymer solution, then placed on a flat surface and irradiated for 1 minute while wet, using a Dymax light system with a 400 watt medium pressure mercury lamp at a distance so that the irradiation intensity is about 2.0 mW / cm ² at 330-340 nm wavelength. The carriers are then washed with deionized water and dried.

Claims (12)

A process for carrying out a solid-phase synthesis comprising the steps of: a) providing a support material having a surface adapted for use in combinatorial syntheses, b) providing a polymer reagent formed by the polymerization of a monomer of the formula

wherein R _{1 is} hydrogen or C ₁ -C ₆ alkyls, and wherein R ₂ and R ₃ , independently of one another, are hydrogen, C ₁ -C ₆ alkyls or phenyls containing one or more reactive substituents selected from

OR ₅ or SR ₅ represents (wherein R _{4 6} alkyl or a heterocyclic ring is a C ₁ -C comprising one or more nitrogen atoms, and R ₅ is a C ₁ -C ₆ alkyl or phenyl containing one or more reactive substituents selected from

c) applying the reagent to the support surface and covalently attaching the polymeric reagent to the support surface d) providing a first reactive monomer selected from nucleotides and amino acids, the monomer comprising a group that is thermochemically reactive with the attached reactive substituent, e) Contacting and reacting the first monomer with the polymer reagent on the support surface under conditions suitable for reacting the associated group with the attached reactive substituent so as to provide a growing polymer chain, and f) providing and subsequently attaching further monomers to the growing one Polymer chain to obtain a desired polymer product. The method of claim 1, wherein the obtained polymer product retain and used in situ. The method of claim 1, wherein the obtained polymer product cut and removed from its position on the carrier. The method of claim 1, wherein the polymer product selected is from the group consisting of polynucleotides, polysaccharides and polypeptides. The method of claim 1, wherein the carrier is provided is in the form of a bead, a wafer, a film, a disk or a plate. The method of claim 5, wherein the carrier comprises: a material selected made of organosilane-treated glass, organosilane-treated silicon, Polypropylene, polyethylene and polystyrene. The method of claim 1, wherein the polymer reagent has the formula:

wherein X = 0-5 mol%, Y = 5-100 mol% and Z = 0-95 mol%. The method of claim 1, wherein the polymer reagent has the formula:

where x = 0-5 mol%, y = 5-100 mol% and z = 0-95 mol%. The method of claim 1, wherein the polymer reagent has the formula:

where m = 15-5 and n = 50-150. The method of claim 1, wherein the polymer reagent has the formula:

where x = 0-5 mol%, y = 5-100 mol% and z = 0-95 mol%. Polymer reagent composition of the formula:

where m = 15-45 and n = 50-150. Polymer reagent composition of the formula:

where x = 0-5 mol%, y = 5-100 mol% and z = 0-95 mol%.